Team:HokkaidoU Japan/Notebook/aggregation protocols

From 2012.igem.org

Aggregation Protocols

Aggregation check

  1. Prepare 2~5 ml LB liquid medium with appropriate antibiotics into round bottom glass tubes that capped with sponge like cap. Plastic (e.g. polypropyrane) tube is not suitable to earn large cluster of cells. And if you used Erlenmeyer flask, prepare 50 ml LB and antibiotics.
  2. Add L-arabinose. Final concentration of L-arabinose becomes 1%.
  3. Suspend colonies into the medium.
  4. Incubate at 34~37C for 24 hrs at 180 rpm. If you wanted to earn large cluster in Erlenmeyer flask, set the temperature at 34C

eCFP and Ag43 expression protocol

  1. Prepare 5 ml LB liquid medium with appropriate antibiotics into round bottom glass tubes that capped with sponge like cap.
  2. Suspend colonies into the medium.
  3. Incubate at 37C for 24 hrs at 180 rpm. For this incubation, pipette 150 ul of this cultivating medium to 96-well microtiter plate every 2 hrs.
  4. Analyze the Absorbance and fluorescence intensity by microtiter plate reader and fluorescence imager.


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